[Abstract ] Objective The purpose of this study was to construct a short hairpin RNA (shRNA) interference lentiviral vector targeting the humanβ-COP gene and to evaluate its inhibitory effect on β-COP in THP-1 cells. Methods We designed and synthesized 4 humanβ-COP-specific oligonucleotide sequences and inserted them into the pGMLV-SC1 vector to construct a recombinant vector fol-lowed by transfection of HEK 293T cells with the recombinant vector and Lenti-HG Mix to produce lentiviruses and detect the viral con-tent.After infecting the THP-1 cells with the packaged lentiviruses , we analyzed the inhibitory effect of β-COP-shRNA on the β-COP gene by quantitative PCR and Western blot . Results Sequencing confirmed that the β-COP-specific oligonucleotide sequences were in-serted into the lentiviral vector and the lentiviruses were packaged in the transfected HEK 293T cells, with the final viral content of 1 × 109 TU/mL.Quantitative PCR showed that the 4 β-COP-shRNA vectors significantly decreased the mRNA expression of β-COP (P<0.01), with interference rates of 16.9 %,32.5%, 74.0%, and 50.3%, respectively.Western blot also indicated their inhibitory effect on the protein expression of β-COP, with an inhibition rate of 76.4% onβ-COP-shRNA3. Conclusion Lentiviral shRNA interference vectors targeting human β-COP were constructed successfully , which could suppress the expression of the human β-COP gene.

AIM: To explore the effect of hypertonic sugar eyes drops with local to the oxygen therapy for severe corneal edema after cataract surgery.METHODS: Totally 68 patients (68 eyes) with severe corneal edema after phacoemulsification were selected from January 2014 to December 2015 in our hospital, who were aged 53-80(64.45±4.24), including 30 males and 38 females.According to different treatment, they were divided into treatment group (34 cases) given conventional therapy + hypertonic sugar eyes drops with local to the oxygen therapy, the control group (34 cases) given conventional treatment + hypertonic sugar eyes drops.Visual acuity and corneal situation were observed during the treatment.RESULTS: Corneal edema fade time of treatment group was 11.62+0.53d, that of control group was 15.23±0.62d, the difference between the two groups was significant (P<0.05).Preoperative best corrected visual acuity (BCVA) and corneal endothelial cell count had no significantly difference between the two groups while the corneal endothelial cell count were significantly different compared with postoperative of the two groups (P<0.05).At 7d after treatment, the BCVA and corneal endothelial cell count had significantly difference between the two groups (P<0.05), which did not have significant difference at 1mo after treatment (P>0.05).The effective rates of the two groups was significantly different (P<0.05).CONCLUSION: Hypertonic sugar eyes drops with local to the oxygen therapy is effective for severe corneal edema after cataract surgery.

This article is based on the Li Dai Zhong Yao Pao Zhi Zi Liao Ji Yao as a clue to review the historical literature and modern data on processing Gentianae Radix et Rhizoma in China, and summarized the relevant contents. Processing methods of Gentianae Radix et Rhizoma have history on the net system, cutting system, heating processing and various accessories processing records, but now the processing methods of Gentianae Radix et Rhizoma are relatively simple, only the methods of net production, ultra-micro-decoction, charcoal, and wine frying are included in the historical "Pharmacopoeia of the People's Republic of China" and local processing specifications. In modern times, the processing methods of wine, licorice gentian, ginger, charcoal, vinegar, and salt for Gentianae Radix et Rhizoma have rationality and quality controllability. Through combing the historical evolution and research progress of processing of Gentianae Radix et Rhizoma, this article provided references for study on modern processing.

Objective To determine the therapeutic effect of recombined rat insulin-like growth factory 1 gene and transforming growth factor beta-1(TGF-?_1)gene on anterior cruciate ligament transection(ACLT)- induced osteoarthritis-like changes in NZW rabbit knee joints.Methods Eighteen NZW rabbits were divided into 3 groups randomly after osteoanhritis was established by ACLT and another six rabbits were used as normal control group(group 1).Chondrocytes which had been transfected with IGF-1 gene,co-transfected with TGF-?_1 and IGF-1 gene(group 3,4)were injected into the rabbits knee joints.Experimental control group(group 2)only had ACLT bul was not transfected.After 4,8 weeks,rabbits were sacrificed and their joints were evaluated by morphological grades,histological examination,in situ hybridization examination,immunohistochemistry exami- nation,and transmission electron microscopy examination(TEM).Results The morphological grades showed that the normal control group had a very significant difference with the experimental control group(P

Objective:Tacrolimus prolonged-release(PR) formulation is a new once-daily formulation of the calcineurin inhibitor tacrolimus,which is currently used in adult liver or kidney transplant patients,and is also gradually widely used in children with nephrotic syndrome.The present study was undertaken to preliminarily investigate the pharmacokinetic characteristics of tacrolimus PR in pediatric nephrotic syndrome recipients.Methods:This single-center open-label prospective study was performed in pediatric nephrotic syndrome recipients.Pharmacokinetic samples were collected from eight pediatric subjects with nephrotic syndrome from Department of Pediatric Nephrology in Peking University First Hospital between June and August 2011.They followed administration of single oral doses of tacrolimus PR formulation at 0.02 mg/kg (n =2),0.05 mg/kg (n =2) and 0.10 mg/kg (n =4).Blood samples were taken before the dose and 1,2,4,6,8,10,12 and 24 h after drug intake.No other medicines or interacting food or drinks were taken during the study period.Blood concentrations were measured using an enzyme multiplied immunoassay technique.Pharmacokinetic analysis was performed using WinNolin Phoenix software Version 6.0 (Pharsight,Cary,NC,USA).Results:The pharmacokinetic data were best described by a non-compartment model.Pharmacokinetic parameters of tacrolimus PR formulation in the 3 ascending doses groups (0.02 mg/kg,0.05 mg/kg and 0.10 mg/kg) were as follows:the maxi mum drug concentrations (Cm=/D) were (1.7 ± 1.0) μg/L,(3.1 ± 1.9) μg/L,(8.0 ± 3.5) μg/L,respectively;Areas under the drug concentration-time curve (AUCo-∞/D) were (47.2 ± 47.1) h · μg/ L,(84.0 ± 13.1) h · μg/L,(175.6 ± 107.1) h · μg/L,respectively;Oral clearance rates were (0.8±0.9) L/(h·kg),(0.4±0.1) L/(h · kg),(1.9 ±1.3) L/(h · kg),respectively;Body weight normalized distribution volumes were (7.0 ± 3.4) L/kg,(12.4 ± 8.4) L/kg and (73.6 ± 68.6) L/kg,respectively.Both mean Cmax normalized level for the administered dose (Cmax/D) and mean AUC0-∞ normalized level for the administered dose (AUC0-∞/D) were higher in the 0.05 mg/kg dosage group than in the 0.02 and 0.10 mg/kg dosage group.There were two peaks in the drug concentrations in every dose group;a primary peak appeared at the end of about 2 h followed by a small secondary peak at h 12,which was more noticeable in the 0.10 mg/kg dose group than in the two lower dosages.Conclusion:The pharmacokinetic characteristics of tacrolimus PR formulation were initially explored in pediatric patients with nephritic syndrome.The data presented form a basis for subsequent larger scale studies on pharmacokinetics of tacrolimus PR formulation in nephritic syndrome children.

BACKGROUNDPeriodontitis and osteoporosis are one of the frequently encountered diseases in post-menopausal women. Estrogen receptors (ERs) regulated bone metabolism. To investigate the possible effect of ER-alpha (α) gene polymorphisms on bone mineral density (BMD) in pre- and post- menopausal Chinese women with chronic periodontitis (CP), we provided sufficient quantitative information concerning the correlation between ER gene polymorphisms and BMD in periodontitis.

METHODSSixty-five post-menopausal and eighty pre-menopausal CP women, and sixty post-menopausal healthy individuals were recruited in this study. Genomic DNA was extracted from oral mucosa swab sample of each subject by the Chelex-100 method. Determination of the ER-α polymorphisms was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique with XbaI and PvuII enzyme. The index for periodontal examination includes clinical attachment loss (CAL) and probing pocket depth (PPD). BMD was measured by dual-energy X-ray absorptiometry (DEXA).

RESULTSThere were no significant differences between the ER-α genotypes of PvuII and XbaI and BMD in post-menopausal and pre-menopausal CP patients, respectively (P >0.05). However, there was association between pre- and post-menopausal CP patients at BMD of lumbar spine L2–L4 (P=0.027) and Ward's BMD (P=0.004). Furthermore, the post-menopausal CP women who carried PvuII TT genotype presented significantly lower Ward's BMD than the pre-menopausal CP women (P=0.007), meanwhile, the post-menopausal CP women who carried XbaI AA genotype presented significantly lower spine L2–L4 BMD than the pre-menopausal CP women (P=0.003).

CONCLUSIONSER-α gene polymorphisms may be a susceptible indicator for BMD variation of lumbar spine L2–L4 and Ward in Chinese pre- and post-menopausal women patients with CP.

Asian Continental Ancestry Group ; genetics ; Bone Density ; genetics ; Chronic Periodontitis ; genetics ; Estrogen Receptor alpha ; genetics ; Female ; Humans ; Polymorphism, Genetic ; genetics ; Postmenopause ; Premenopause

Taking guizhi fuling capsule (GZFL) for instance, a new method about reference Chinese medicine preparation which was used as standard substance for the quality evaluation of complex Chinese medicine preparation by the fingerprint of reference preparation instead of standard fingerprint was proposed. It could eliminate the errors from different instruments, chromatographic columns and solve the problem of similarity matching in the absence of standard fingerprint. The qualification of reference GZFL was evaluated according to the quality control method of GZFL from Chinese Pharmacopoeia. Then multiple batches of GZFL were estimated, taking fingerprint of reference preparation and standard fingerprint as references, respectively, at different instruments and chromatographic columns. Finally, the packaging and expiration date for reference GZFL were confirmed according to the results of stability investigation. The results indicated that the fingerprint of reference GZFL could be used to assess the quality of GZFL better than standard fingerprint. The data of accelerated stability and long-term stability test demonstrated that reference GZFL was stable in the conditions of double blister package. Therefore, reference GZFL can be used as standard substance in quality control of GZFL.
Capsules ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; standards ; Reference Standards

OBJECTIVETo explore the better therapy in the treatment of ganglion.

METHODSNinety cases of ganglion were randomized into a two-way quintuple puncture group, a common quintuple puncture group and a fire needling group, 30 cases in each one. In the two-way quintuple puncture group, the "9-in-1" multiple penetrating needling technique was used. In the common quintuple puncture group, the traditional "5-in-1" multiple penetrating needling technique was applied. In the fire needling group, the traditional multiple fire needling technique was adopted. The treatment was given once a day, 3 treatments made one session and the efficacy was analyzed statistically after 1 session treatment in the three groups.

RESULTSAll of the three therapeutic methods achieved the efficacy on ganglion. The curative rate was 96. 7% (29/30) in the two-way quintuple puncture group, which was better obviously than 66.7% (20/30) in the common quintuple puncture group and 60. 0% (18/30) in the fire needling group (both P<0. 01).

CONCLUSIONThe two-way quintuple puncture technique achieves the remarkably superior efficacy on ganglion as compared with the common quintuple puncture technique and fire needling technique.

Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Female ; Ganglion Cysts ; therapy ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

BACKGROUNDTo establish a new assay for detecting the quantity of HBV DNA with PCR and enzyme-linked immunosorbent assay(ELISA).

METHODSThe products of PCR using primers pre-labeled with biotin were hybridized with the capture probes that were immobilized on the microtiter strips and then bond with Sav-Ap. The quantity of DNA was detected by measuring the yellow color at 405 nm wave length.

RESULTSTotally 125 sera from patients with hepatitis B were tested for HBV DNA by this method,the sera were also tested for HBV immunological markers by solid phase radio immuno-assay (SPRIA). The HBV positive rate with PCR-ELISA was 64.9% (24/37) in samples which were positive for HBsAg, HBeAg and HBcAb; and 34.2% (13/38) in sera which were positive for HBsAg, HBeAb and HBcAb; in sera positive for HBsAg and HBcAb or only HBcAb, the positive rate was 6.7% (1/15) and 5.9% (2/34) respectively.

CONCLUSIONSThe PCR ELISA assay is simple and suitable for clinical laboratory in quantitative determination of HBV DNA.

DNA Probes ; DNA, Viral ; analysis ; biosynthesis ; genetics ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B ; virology ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Polymerase Chain Reaction ; Sensitivity and Specificity